超高效液相色谱-三重四极杆质谱法检测柴胡中藏柴胡的掺伪  

Determination of adulteration of Bupleurum marginatum var.stenophyllum in Bupleurum by UPLC-QqQ-MS

作　　者：李敏 李婷 韩潇潇 马潇 李冬华 张平 杨玲霞 LI Min;LI Ting;HAN Xiao-xiao;MA Xiao;LI Dong-hua;ZHANG Ping;YANG Ling-xia(Gansu Institute for Drug Control,Lanzhou 730030,China;Center for Inspection of GSMPA,Lanzhou 730030,China;Beijing Sunflower Pharmaceutical Research Co.,Ltd.,Beijing 100050,China)

机构地区：[1]甘肃省药品检验研究院,兰州730030 [2]甘肃省药品监督管理局审核查验中心,兰州730030 [3]北京葵花医药研究有限责任公司,北京100050

出　　处：《药物分析杂志》2024年第6期999-1008,共10页Chinese Journal of Pharmaceutical Analysis

基　　金：甘肃省药品监督管理局,中药材及饮片质量控制重点实验室(2023GSMPA-KL06);甘肃省药品监督管理局,甘肃省中药饮片炮制规范化研究(2022GSMPA-0075);国家药监局中药材及饮片质量控制重点实验室项目(2021GSMPA-KL04,2022GSMPA-KL12,2022GSMPA-KL10,2020GSMPA-KL15)。

摘　　要：目的:建立检测柴胡及藏柴胡中柴胡皂苷K成分的液相色谱-质谱法(LC-MS)检查方法,用以快速鉴别北柴胡是否为藏柴胡或混有藏柴胡。同时对16批藏柴胡和157批柴胡饮片中的柴胡皂苷K进行检测。方法:采用超高效液相色谱-三重四极杆串联质谱(UPLC-QqQ-MS)对柴胡及藏柴胡中柴胡皂苷K进行检测,色谱柱为Phenomenex Kinetex C18(100 mm×2.1 mm, 1.7μm),流动相为乙腈(A)-水(B),梯度洗脱(0~5 min, 15%A→25%A;5~30 min, 25%A→30%A;30~35 min, 30%A→90%A;35~36 min, 90%A→15%A;36~40 min, 15%A),流速0.3 mL·min^(-1),柱温25℃,进样量5μL;采用电喷雾离子源(ESI-),负离子模式扫描,多反应监测(MRM)模式检测,选择柴胡皂苷K特征分子离子峰m/z 943.5→797.3、m/z 943.5→635.3和m/z 943.5→781.3作为检测离子对。结果:16批藏柴胡中柴胡皂苷K的量远高于限度要求,157批柴胡饮片中有8批样品中均检出藏柴胡类成分柴胡皂苷K。结论:所建立的方法经方法学验证,专属性强,以期制定柴胡中藏柴胡成分检查项补充检验方法,提升柴胡饮片质量控制标准及真伪鉴别研究,可进一步有效控制其质量,保障临床疗效。Objective:To establish a liquid chromatography mass spectrometry(LC-MS)method for the detection of nepasaikosaponin K in Bupleurum marginatum var.stenophyllum,which could be used to quickly identify whether Bupleurum marginatum var.stenophyllum was substituted for Bupleurum or was mixed with Bupleurum.At the same time,nepasaikosaponin K was detected in 16 batches of Bupleurum marginatum var.stenophyllum and 157 batches of bupleurum decoction pieces.Methods:Ultra-high performance liquid chromatography(UPLC)coupled with triple quadruple mass spectrometry(QqQ MS)was used to detect nepasaikosaponin K in Bupleurum and Bupleurum marginatum var.stenophyllum.Determination was carried out with the application of a Phenomenex Kinetex C18(2.1 mm×100 mm,1.7μm)column at temperature of 25℃.The mobile phase was composed of acetonitrile(A)and water(B)with gradient elution(0-5 min 15%A→25%A;5-0 min,25%A→30%A;30-35 min,30%A→90%A;35-36 min,90%A→15%A;36-40 min,15%A)at a flow rate of 0.3 mL·min^(-1).The electrospray ionization(ESI-)source was performed in multiple reaction monitoring(MRM)mode of the transitions of m/z 943.5→797.3,m/z 943.5→635.3,and m/z 943.5→781.3.Results:The contents of nepasaikosaponin K detected in 16 batches of Bupleurum marginatum var.stenophyllum were much higher than minimum requirements.Nepasaikosaponin K was detected in 8 out of 157 batches of Bupleurum.Conclusion:The established method is verified by methodology to be specific,so as to develop a supplementary test method for the components of Bupleurum and Bupleurum marginatum var.stenophyllum.to improve the quality control standard and authentic identification investigation,and to effectively control the quality of Bupleurum decoction pieces and guarantee its clinical efficacy.

关 键 词：柴胡 藏柴胡 柴胡皂苷K 补充检验方法 超高液相色谱-三重四极杆串联质谱 真伪鉴别 质量问题 

分 类 号：R917[医药卫生—药物分析学]